Abstract 16002: A 3D Engineered Model of Human Induced Pluripotent Stem Cell Derived Cardiomyocytes Reveals Functional and Structural Characteristics of β-Myosin Heavy Chain Mutation Mediated Hypertrophic Cardiomyopathy
Introduction: Hypertrophic Cardiomyopathy (HCM) affects 1 in 500 people and may result in heart failure, stroke and sudden cardiac death. However, mechanism based research has been limited by the poor availability of viable human HCM cardiomyocytes for in vitro study. A source of functionally and structurally mature human HCM cardiomyocytes is necessary to advance our understanding of this disease and for development of new treatments.
Hypothesis: Induced Pluripotent Stem Cell Derived Cardiomyocytes (iPSC-CMs) and self-assembled 3D microtissues generated for an HCM patient with an inherited mutation in the MYH7 gene (R403Q) will recapitulate the structural, cellular and electrical HCM phenotype of the patient.
Methods and Results: Control or HCM patient vector-free iPSCs were generated using episomal reprogramming. CMs were generated using small molecule based manipulation of the Wnt signaling pathway. We utilized flexible two-post 3D microtissue technologies as a mechanism of mechanotransductive maturation of hiPSC-CMs. 3D microtissues improve cellular alignment, sarcomere organization and electrophysiological parameters such as action potential duration (1Hz pacing APD80 of 2D monolayers=526.8±22.2 vs. 395±8.7 ms in 3D, p<0.05) compared to 2D monolayers. HCM 3D microtissue formation occurred at a significantly slower rate than controls and once formed the 3D HCM microtissues were larger than controls 2.89 ± 0.31 mm2 vs 1.70 ± 0.17 mm2 (p<0.05). In control experiments adenoviral mediated gene transfer of R403Q MYH7 in normal control hiPSC-CMs produced similar results to the HCM patient specific cell line. HCM 3D microtissues had poor directional alignment, sarcomere organization and intercellular connectivity and a greater propensity for arrhythmias (47% vs 9%). In our model, HCM deficits in 3D cellular organization led to a significant decrease in total force generation of the tissue (1.73 ± 0.27 vs. 0.14 ± 0.04 mN, p<0.05).
Conclusions: Self-assembly of human iPSC-CM 3D cardiac microtissues serves as an important method for hiPSC-CM maturation and study of structural heart disease. We have developed a human in vitro 3D tissue model of a patient’s R403Q HCM which recapitulates the electrical, structural and organization hallmarks of HCM.
- Biology, developmental
- Hypertrophic cardiomyopathy
- Cardiac development
- Cardiac hypertrophy
- Tissue engineering
Author Disclosures: K. Campbell: None. J. Davis: None. A. Monteiro da Rocha: None. G. Guerrero-Serna: None. X. Li: None. L. Mundada: None. E. Hoopingarner: None. A. Helms: None. S. Day: None. J. Jalife: None. J. Fu: None. T.J. Herron: None.
- © 2016 by American Heart Association, Inc.